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Related Experiment Video

Updated: Jan 18, 2026

Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy 3D-SIM
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A correlation-based tool for quantifying membrane periodic skeleton associated periodicity.

Sam K Vanspauwen1, Virginia Luque-Fernández1, Hanne B Rasmussen1

  • 1Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Frontiers in Neuroinformatics
|September 8, 2025
PubMed
Summary

We developed Napari-WaveBreaker, an open-source tool to quantify the periodic localization of proteins within the membrane-associated periodic skeleton (MPS). This method accurately analyzes MPS periodicity and co-distribution, aiding in understanding neuronal structure.

Keywords:
autocorrelationaxon initial segmentcross-correlationmembrane-associated periodic skeletonnaparisoftwaresuper-resolution microscopy

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biophysics

Background:

  • Super-resolution microscopy revealed the membrane-associated periodic skeleton (MPS), a neuronal cytoskeletal structure with actin rings spaced 190 nm apart.
  • Numerous proteins associate with the MPS, but tools for quantifying their periodic localization are limited.

Purpose of the Study:

  • To develop an open-source tool for accurate and unbiased quantification of MPS periodicity and protein co-distribution.
  • To provide a framework for analyzing molecular organization within the MPS.

Main Methods:

  • Development of Napari-WaveBreaker, an open-source plugin for the Napari image viewer.
  • Utilized autocorrelation to quantify MPS periodicity and cross-correlation for assessing co-distribution of targets.
  • Evaluated performance using simulated datasets and STED microscopy images.

Main Results:

  • Napari-WaveBreaker accurately quantified MPS periodicity and detected spatial shifts between periodic targets.
  • The tool demonstrated robustness across varying image qualities.
  • Successfully distinguished between periodic and non-periodic protein distributions.

Conclusions:

  • Napari-WaveBreaker offers an unbiased, quantitative framework for analyzing MPS periodicity and co-distribution.
  • Enables new insights into the molecular organization and modulation of the MPS.